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ID   PFL260     preliminary; circular DNA; SYN; 3642 BP.
XX
AC   M29362;
XX
DT   16-JAN-1991 (Rel. 6, Created)
DT   01-JUL-1995 (Rel. 12, Last updated, Version 1)
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DE   E. coli plasmid vector pFL260 - complete.
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KW   cloning vector.
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OS   Cloning vector
OC   Artificial sequences; Cloning vehicles.
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RN   [1]
RP   1-3642
RC   pFL200 from pFL114
RC   pFL236 from pFL200 & pFL166
RC   pFL245 from pFL200
RC   pFL257 from pFL236
RC   pFL260 from pFL245
RC   pFL261 from pFL257
RA   Larimer F.W., Mural R.J., Soper T.S.;
RT   "Versatile protein engineering vectors for mutagenesis,
RT   expression and hybrid enzyme formation";
RL   Protein Engineering 3:227-231(1990).
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RN   [2]
RC   pFL260
RA   Larimer F.W.;
RT   ;
RL   Submitted (23-OCT-1989) to GenBank on computer-readable media by:
RL   Larimer F.W., ORNL, PO Box 2009, Oak Ridge TN 37831, USA.
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RN   [3]
RC   pFL114 from pBR322 & rbc gene
RC   pFL113 from pBR322 & pFL159, rbc gene
RC   pFL161 from pACYC184 & pFL159, rbc gene
RC   pFL166 from pACYC184 & rbc gene
RC   pFL167 from pFL113 & pFL161
RC   NOTE: see J. Biol. Chem. 263:2575-2575(1988)
RA   Larimer F.W., Lee E.H., Mural R.J., Soper T.S., Hartman F.C.;
RT   "Intersubunit location of the active site of ribulose-bisphosphate
RT   carboxylase/oxygenase as determined by in vivo hybridization of
RT   site-directed mutants";
RL   J. Biol. Chem. 262:15327-15329(1987).
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RN   [4]
RC   pGL101B from pGL101 & linker
RC   pFL23 from pRR36 & pGL101B
RC   pFL34 from pRR36 & pDR540
RC   pFL25 from pRR36 & linker & pFL23
RC   pDR540em from pDR540 & ery gene
RC   pFL35 from pFL25 & pDR540em
RA   Larimer F.W., Machanoff R., Hartman F.C.;
RT   "A reconstruction of the gene for ribulose bisphosphate
RT   carboxylase from Rhodospirillum rubrum that expresses the
RT   authentic enzyme in Escherichia coli";
RL   Gene 41:113-120(1986).
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RN   [5]
RC   pRR2119 from Rhodospirillum rubrum rbc gene
RC   pRR36 from pRR2119
RA   Somerville C.R., Somerville S.C.;
RT   "Cloning and expression of the Rhodospirillum rubrum
RT   ribulose-bisphosphate carboxylase gene in E. coli";
RL   Mol. Gen. Genet. 193:214-219(1984).
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RN   [6]
RC   pFL114 from pBR322 & rbc gene
RC   pFL159 from pBR322 & rbc gene & tac promoter
RA   Hartman F.C., Soper T.S., Niyogi S.K., Mural R.J., Foote R.S.,
RA   Mitra S., Lee E.H., Machanoff R., Larimer F.W.;
RT   "Function of Lys-166 of Rhodospirillum rubrum ribulosebisphosphate
RT   carboxylase/oxygenase as examined by site-directed mutagenesis";
RL   J. Biol. Chem. 262:3496-3501(1987).
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CC   NM (pFL260)
CC   CM (yes)
CC   NA (ds-DNA)
CC   TP (circular)
CC   ST ()
CC   TY (plasmid)
CC   SP ()
CC   HO (E.coli)
CC   CP ()
CC   FN (mutagenesis)(expression)(fusion)
CC   SE ()
CC   PA (ColE1)(pFL34)(pFL245)
CC   BR (pFL261)
CC   OF ()
CC   OR ()
XX
FH   Key             Location/Qualifiers
FH
FT   misc_feature    0..0
FT                   /note="1. pBR322 remove HindIII-BamHI 346bp 30..376,
FT                   \ 4015bp
FT                   2. E.coli HindIII-BamHI, tac prom/rbc gene
FT                   -> pFL114 5000bp
FT                   1. pFL114 remove AatII-EcoRI 69bp 4291..4360, 5000bp
FT                   2. pBS+ AatII-EcoRI 946bp 3140..3204..882, f1 ori
FT                   -> pFL200 6000bp
FT                   1. pFL200 remove 17bp aggaaacaggatccatg,
FT                   \ RBS-ATG/6000bp
FT                   2. oligo 17bp aggaggaaggatccatg, RBS-ATG
FT                   -> pFL245 6000bp
FT                   1. pFL245 remove NcoI-EagI,
FT                   \ rbc gene/pBR322 EagI 940/3642bp
FT                   -> pFL260 3642bp"
FT   promoter        318..345
FT                   /note="PRO E. coli tac (trp and lac)"
FT   RBS             380..384
FT                   /note="RBS"
FT   rep_origin      942..942
FT                   /note="ORI E. coli pMB1 (ColE1 and pBR322)"
FT   CDS             complement(1702..2564)
FT                   /note="ANT E. coli beta-lactamase gene (bla)
FT                   ampicillin resistance gene (apr/amp)"
XX
SQ   Sequence 3642 BP; 924 A; 906 C; 963 G; 849 T; 0 other;
     gaattctggc gaatcctctg accagccaga aaacgacctt tctgtggtga aaccggatgc
     tgcaattcag agcgccagca agtgggggac agcagaagac ctgaccgccg cagagtggat
     gtttgacatg gtgaagacta tcgcaccatc agccagaaaa ccgaattttg ctgggtgggc
     taacgatatc cgcctgatgc gtgaacgtga cggacgtaac caccgcgaca tgtgtgtgct
     gttccgctgg gcatgccagg acaacttctg gtccggtaac gtgctgagcc cggccaagct
     tactccccat ccccctgttg acaattaatc atcggctcgt ataatgtgtg gaattgtgag
     cggataacaa tttcacacag gaggaaggat ccatgggtcg accggccggt caaagcagaa
     gacgcggagg ctctcccctc cgcgtctttt tttgagcttt ttcgatgggg atccgctgcc
     tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
     cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     ttggcgggtg tcggggcgca gccatgaccc agtcacgtag cgatagcgga gtgtatactg
     gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc ggtgtgaaat
     accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcaatg
     ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     gccagttaat agtttgcgca acgttgttgc cattgctgca ggcatcgtgg tgtcacgctc
     gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaaca cgggataata ccgcgccaca
     tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata
     ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta
     gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta
     agaaaccatt attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg
     tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt
     cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg
     tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt
     gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggaa
     attgtaaacg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat cagctcattt
     tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata gaccgagata
     gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt ggactccaac
     gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc atcaccctaa
     tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa agggagcccc
     cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg gaagaaagcg
     aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt aaccaccaca
     cccgccgcgc ttaatgcgcc gctacagggc gcgtcgcgcc attcgccatt caggctgcgc
     aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     aaaacgacgg ccagtgaatt gtaatacgac tcactatagg gc
//